百合LbKCS5和LbKCS17基因的克隆与表达分析
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引用本文:赵子贤,刘 立,谢 天,阎俊卉,文锦芬.百合LbKCS5和LbKCS17基因的克隆与表达分析[J].西北植物学报,2022,42(8):1273~1281
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作者单位
赵子贤,刘 立,谢 天,阎俊卉,文锦芬* (昆明理工大学 建筑与城市规划学院昆明 650500) 
基金项目:国家自然科学基金(31760588);
中文摘要:β 酮脂酰辅酶A合成酶(KCS)是超长饱和脂肪酸链(VLCFAs)生物合成中的限速酶,该研究以百合(Lilium brownii var. viridulum Baker.)‘黄天霸’cDNA为模版, 采用RT PCR方法克隆了LbKCS5和LbKCS17基因的CDS序列,并进行序列分析,采用qRT PCR方法分析其基因表达以及胁迫诱导特征。结果表明:(1)LbKCS5和LbKCS17分别包含689和1 238 bp完全阅读框(ORF), 编码186和291个氨基酸;进化分析显示, LbKCS5与油棕KCS5、LbKCS17与莲KCS17序列相似性最高,分别为82.61%和77.62%。(2)基因表达分析结果显示, LbKCS5和LbKCS17 在百合营养器官中均有表达,二者在叶中表达量最高,在根中最少;在花器官中,LbKCS5的表达量在雄蕊中最高,LbKCS17在花药中最高,在花瓣中二者的表达量最低;在花蕾生长发育过程中,花瓣中2个基因的表达均先升高后下降,并于黄蕾期达到峰值;叶中LbKCS5表达量在黄蕾期达到最高,而LbKCS17在叶中的表达整体较低。(3)失水胁迫能提高这2个基因的转录表达,低温诱导LbKCS5表达量的增加,但却降低了LbKCS17的表达。研究表明,LbKCS5和LbKCS17在百合不同器官中均有表达,且均响应失水和低温胁迫,这为研究百合耐失水和低温胁迫以及新品种选育提供了理论支持。
中文关键词:LbKCS  百合  蜡质  基因克隆  表达分析
 
Cloning and Expression Analysis of LbKCS5 and LbKCS17 in Lilium brownii var. viridulum Baker
Abstract:β ketoacyl CoA synthase (KCS) is the rate limiting enzyme in the biosynthesis of very long chain fatty acids (VLCFAs). In this study, Lilium brownii var. viridulum Baker ‘Huangtianba’ was used as material, the CDS sequences of LbKCS5 and LbKCS17 genes were cloned by RT PCR using cDNA as a template, and sequence analysis, qRT PCR were performed to study their gene expression and stress induction analysis. The results showed that: (1) LbKCS5 and LbKCS17 contained 689 bp and 1 238 bp complete reading frames (ORFs), encoding 186 and 291 amino acids, respectively. Sequence similarity between LbKCS5 and oil palm KCS5,LbKCS17 and lotus KCS17 were 82.61% and 77.62%, respectively. (2) Expression level analysis indicated that both LbKCS5 and LbKCS17 were expressed in the vegetative organs of lily, with the highest expression in leaves and the least in roots. In floral organs, petals had the two genes’ lowest expression, the highest expression levels of LbKCS5 and LbKCS17 were in stamens and anthers, respectively. During the growth and development of flower buds, the two genes’ expression increased first and then decreased, reaching a peak at the yellow bud stage in petals; in leaves LbKCS5 reached the highest at the yellow bud, while the expression of LbKCS17 was generally low. (3) The transcriptional expression levels of these two genes surged under dehydration stress, and low temperature can induce LbKCS5 expression rinsing, but reduce the expression of LbKCS17; Our results show that LbKCS5 and LbKCS17 were expressed in different organs of lily and responded to dehydration and low temperature stresses, which provided a feasible approach and theoretical supporting for future studies on improving these two stresses tolerance and breeding of new varieties of lily.
keywords:LbKCS  lily  wax  gene cloning  expression analysis
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